The alphaviruses: gene expression, replication, and evolution.

The crystallographically determined structure of a soluble fragment from the major envelope protein of a flavivirus reveals an unusual architecture. The flat, elongated dimer extends in a direction that would be parallel to the viral membrane. Residues that influence binding of monoclonal antibodies lie on the outward-facing surface of the protein. The clustering of mutations that affect virulence in various flaviviruses indicates a possible receptor binding site and, together with other mutational and biochemical data, suggests a picture for the fusion-activating, conformational change triggered by low pH.

The envelope glycoprotein from tick-borne encephalitis virus at 2 Å resolution

Present and future arboviral threats.

Dengue virus enters a host cell when the viral envelope glycoprotein, E, binds to a receptor and responds by conformational rearrangement to the reduced pH of an endosome. The conformational change induces fusion of viral and host-cell membranes. A three-dimensional structure of the soluble E ectodomain (sE) in its trimeric, postfusion state reveals striking differences from the dimeric, prefusion form. The elongated trimer bears three ‘fusion loops’ at one end, to insert into the host-cell membrane. Their structure allows us to model directly how these fusion loops interact with a lipid bilayer. The protein folds back on itself, directing its carboxy terminus towards the fusion loops. We propose a fusion mechanism driven by essentially irreversible conformational changes in E and facilitated by fusion-loop insertion into the outer bilayer leaflet. Specific features of the folded-back structure suggest strategies for inhibiting flavivirus entry.

Structure of the dengue virus envelope protein after membrane fusion

Arthropod-borne viruses (arboviruses) are the causative agents of some of the most important emerging infectious diseases and are responsible for significant global public health problems (Gubler, 2001). Of these viruses, the genus Alphavirus belongs in the family Togaviridae, and the species in this genus cause diseases ranging from mild febrile illness to severe polyarthritis to encephalitis. One alphavirus, chikungunya virus (CHIKV), caused recent outbreaks associated with severe morbidity. Traditionally, CHIKV causes a febrile illness similar to that seen in dengue virus infections. The hallmark feature of CHIKV disease is a debilitating and prolonged arthralgic syndrome that primarily affects the peripheral small joints. While the acute febrile phase of the illness normally resolves within a few days, the pain associated with CHIKV infection of the joints typically persists for weeks or months causing serious economic and social impact on both the individual and the affected communities. During the 2005–2007 explosive epidemics on the Indian Ocean islands and in India, anecdotal cases of CHIKV-associated deaths, encephalitis and neonatal infections were reported. Here, we review outbreaks of CHIKV disease and present a comprehensive analysis and comparison of clinical presentation, virus– vector associations and viral genetics to assess the possibility of further expansion and re-emergence of CHIKV illness.

Changing patterns of chikungunya virus: re-emergence of a zoonotic arbovirus

Strains of myxoma virus associated with a case-mortality rate in wild rabbits of 90—95% can be distinguished from those associated with a case-mortality rate of more than 99% by the use of a standardized virulence test carried out in groups of five laboratory rabbits, and tests carried out on laboratory rabbits are closely correlated with the results obtained with Australian wild rabbits. The test, which attempts to mimic natural mosquito transmission, consists of the intradermal inoculation in one site of about 5 rabbit-infectious doses of the virus, and the subsequent observation of the rate of progress of clinical symptoms and ultimate fate of the rabbit. The mean survival time provides a figure which allows satisfactory classification of all known myxoma virus strains into one of four grades of virulence, very attenuated strains being recognized by their low case-mortality rates.Using this test, 92 strains of myxoma virus have been compared. These comprised 6 laboratory strains, 62 strains recovered from Australian wild rabbits or from wild caught mosquitoes, between February, 1951 and March, 1955, 19 strains recovered from naturally infected rabbits in Europe between October, 1953 and August, 1955, 3 strains from South America, and 2 from California. Eight types have been described and illustrated, differentiation being based on their virulence and the clinical picture of the disease they cause in laboratory rabbits. The differences are summarized in Text-fig. 2. Four of the virus types produce recognizably different pocks on the chorioallantois of developing chick embryos.In Australia the repeatedly introduced standard laboratory strain of virus has been replaced by a moderately virulent strain, which appears to be better adapted for survival than either the highly virulent standard laboratory strain, or less virulent variants of which only a few had been recovered up to 1955. Strains of the type dominant in 1953–55 have appeared independently in many widely separated parts of Australia. Examination of these attenuated variants by pure clone techniques showed that the variability in symptomatology associated with them is not due to a mixture of virus strains, but to the expression of differences in innate resistance of the host animals which are obscured by the overwhelming virulence of the highly virulent strains.Only one introduction of a different strain of virus was made in Europe. All field strains recovered during the first two years after the introduction were of high virulence, but a few attenuated strains appeared in 1955, and they have since become widespread. One of them appears to be a relatively stable mixture of a highly virulent and a greatly attenuated strain.The collection of the large numbers of strains of virus described in this paper has been made possible only by the generous collaboration of many people. In Australia we would like to acknowledge the great help afforded us by Mr F. N. Ratcliffe, officer-in-charge of the Wildlife Survey Section, Commonwealth Scientific and Industrial Research Organization and his officers, especially Messrs J. H. Calaby, G. Douglas, A. L. Dyce, B. V. Fennessy, E. W. Lines, K. Myers, R. Mykytowycz, W. E. Poole and E. Waterhouse. Messrs B. D. Robinson and T. Pearce of the Lands Department of Victoria have also provided us with specimens, as have Mr G. Edgar, Director of the Veterinary Research Station, Glenfield, New South Wales, Dr D. Surrey Dane of the Institute of Medical and Veterinary Science, Adelaide, Mr D. Gooding, Department of Agriculture, Western Australia, and Messrs T. B. Alexander and A. F. Ryan, Department of Agriculture, Tasmania.We are indebted to Dr H. de B. Aragao of the Instituto Oswaldo Cruz, Rio de Janeiro, and Dr B. Szyfres, of the Laboratorio de Biologia Animal, Uruguay, for specimens from South America; Dr D. G. McKercher of the University of California School of Veterinary Medicine, Davis, California, for the Californian strains; Drs P. Lepine and H. Jacotot, of the Institut Pasteur, Paris, for strains from France; and Mr J. R. Hudson, of the Veterinary Laboratory, Weybridge, and Dr C. H. Andrewes of the National Institute for Medical Research, Mill Hill, for material from England.We are grateful to Dr S. Fazekas de St Groth and Dr G. S. Watson of this University for advice on the statistical treatment of the data.Mr V. Paral took the photographs.

A comparison of the virulence for European rabbits (Oryctolagus cuniculus) of strains of myxoma virus recovered in the field in Australia, Europe and America.

Host cell selection of murray valley encephalitis virus variants altered at an RGD sequence in the envelope protein and in mouse virulence

Annually for 4 years groups of young Australian wild rabbits have been captured during non-epizootic periods from areas in which myxomatosis has occurred during the previous summer. The intensity of the preceding epizootic was measured by testing a sample of survivors for antibody. The captured young rabbits were raised in the laboratory until they were about 4 months old and then inoculated intradermally with small doses of the slightly attenuated myxoma virus strain Aust/Corowa/12–52/2 (KM 13), all samples of virus being derived from the same batch which has been stored at -70°C.The results show there is a significant negative correlation between the mortality rate and the degree of exposure to myxomatosis of the forbears of the tested animals. Passive and active immunization have been excluded and this result is ascribed to increased genetic resistance.We are greatly indebted to Mr. F. N. Ratcliffe, Officer in Charge of the Wildlife Survey Section of the Commonwealth Scientific and Industrial Organization, and to his officers, especially Messrs W. E. Poole, K. E. Myers, G. Douglas, B. V. Fennessy, H. Frith and K. Keith for their invaluable assistance in collecting rabbits and serum samples.We have had valuable discussions with Dr W. R. Sobey, Dr P. J. Claringbold and Dr G. S. Watson on genetical and statistical aspects of the problems under study.

/pdf/studies-in-the-epidemiology-of-infectious-myxomatosis-of-2raip1iqat.pdf

Studies in the epidemiology of infectious myxomatosis of rabbits. V. Changes in the innate resistance of Australian wild rabbits exposed to myxomatosis.

Ross River virus (RRV) is an Australian alphavirus that is often responsible for chronic epidemic polyarthritis and myalgia in humans. Past studies have shown severe disruption of striated muscle fibers to be prominent in RRV pathology in mice; in the present study, macrophages were directly implicated as the primary mediators of muscle damage. General immunosuppressive therapies had only minor effects on mortality and morbidity in RRV-infected mice, with no inhibition of muscle damage. Treatment of mice with macrophage-toxic agents (e.g., silica) prior to RRV infection completely abrogated disease symptoms without significantly affecting titers of virus in organs. Further studies found that clinical signs of infection and muscle damage correlated with a massive influx of macrophages into hind leg muscle, whereas no such infiltrate or damage was observed for silica-treated mice. These observations are significant for the human disease context, as monocytic cells have been detected in the synovial effusions of persons with epidemic polyarthritis.

https://www.jstor.org/stable/info/30111270

Macrophage-Induced Muscle Pathology Results in Morbidity and Mortality for Ross River Virus-Infected Mice

Pools of mosquitoes collected in the Murray Valley in February, 1974, during an encephalitis epidemic yielded 239 isolates of 11 distinct viruses. These included 39 isolates of MVE virus, an incriminated causative agent of encephalitis in man, and 111 isolates of Kunjin virus, a probable causative agent. An additional isolate of MVE virus was recovered from the serum of a white-faced heron, Ardea novaehollandiae. The other 9 viruses comprised the Alpha-viruses Ross River and Sindbis, the Flavivirus, Edge Hill, the Bunyaviruses, Koongol and Wongal, the undefined Kowanyama virus, and 3 previously undescribed viruses. One isolate of MVE virus was recovered from a total of 180 Culex australicus, and all other isolates of all viruses were from Culex annulirostris. The combined apparent infection rate of MVE and Kunjin viruses in Cx annulirostris was 1/913. The concurrent activity of so many seemingly transient arboviruses, their wide geographical distribution and temporal considerations all weaken the widely held hypothesis that during the pre-epidemic period MVE virus is introduced into the Murray Valley by viraemic waterbirds from distant areas of tropical enzooticity.

Viruses recovered from mosquitoes and wildlife serum collected in the Murray Valley of South-eastern Australia, February 1974, during an epidemic of encephalitis

Ian D. Marshall

Papers

A comparison of the virulence for European rabbits (Oryctolagus cuniculus) of strains of myxoma virus recovered in the field in Australia, Europe and America.

Host cell selection of murray valley encephalitis virus variants altered at an RGD sequence in the envelope protein and in mouse virulence

Studies in the epidemiology of infectious myxomatosis of rabbits. V. Changes in the innate resistance of Australian wild rabbits exposed to myxomatosis.

Macrophage-Induced Muscle Pathology Results in Morbidity and Mortality for Ross River Virus-Infected Mice

Viruses recovered from mosquitoes and wildlife serum collected in the Murray Valley of South-eastern Australia, February 1974, during an epidemic of encephalitis